TABLE 1.

Detail of the primers used in optimization of the PMAxx^TM -based qPCR assay.

Gene symbol	Primer sequence (5′-3′)	Accession no.	Product size (bp)	Annealing temperature (°C)	Amplification efficiency (%)
invA	F: AAACCTAAAACCAGCAAAGG R: TGTACCGTGGCATGTCTGAG	M90846.1	605	59	92
invA	F: TGGGGCGGAATATCATGACG R: AGGAAGGTACTGCCAGAGGT	M90846.1	1045	60	80
ompA	F: TACGCTGGTGCTAAACTGGGCT R: AGCGCGAGGTTTCACGTTGTCA	NC_003197.2	882	60	83
ompA	F: CAGTACCATGACACCGGCTT R: ATTCCAGACGGGTTGCGATT	NC_003197.2	385	60	90

Specificity of each primer pairs was confirmed through melting curve analysis and gel electrophoresis. Sequence of the invA 605 bp was sourced from literature (Akiba et al., 2011). Among the listed primers in Table 1, invA having 605 bp sequence length had higher amplification efficiency and target specificity. Therefore, primer pair labeled as invA 605 bp was used in the quantification of viable and non-viable load of Salmonella Enteritidis and Salmonella Typhimurium through PMAxx^TM -based qPCR from soil samples incubated at 5, 25, and 37°C for up to 6 weeks.